← Product Code [CFR](/submissions/CH/subpart-b%E2%80%94clinical-chemistry-test-systems/CFR) · K220134

# cobas pure integrated solutions, Glucose HK Gen.3, ISE indirect Na for Gen.2, ONLINE DAT Methadone II, Elecsys TSH (K220134)

_Roche Diagnostics · CFR · Sep 16, 2022 · Clinical Chemistry · SESE_

**Canonical URL:** https://fda.innolitics.com/submissions/CH/subpart-b%E2%80%94clinical-chemistry-test-systems/CFR/K220134

## Device Facts

- **Applicant:** Roche Diagnostics
- **Product Code:** [CFR](/submissions/CH/subpart-b%E2%80%94clinical-chemistry-test-systems/CFR.md)
- **Decision Date:** Sep 16, 2022
- **Decision:** SESE
- **Submission Type:** Traditional
- **Regulation:** 21 CFR 862.1345
- **Device Class:** Class 2
- **Review Panel:** Clinical Chemistry
- **Attributes:** Pediatric

## Indications for Use

cobas pure integrated solutions is an automated analyzer, intended for running qualitative, semiguantitative and quantitative clinical chemistry and immunochemistry assays as well as ion selective measurements. Glucose HK Gen.3 is an in vitro test for the quantitative determination of glucose in human serum, plasma, urine and CSF on Roche/Hitachi cobas c systems. Glucose measurements are used in the diagnosis and treatment of carbohydrate metabolism disorders including diabetes mellitus, neonatal hypoqlycemia, idiopathic hypoglycemia and pancreatic islet cell tumors. The ISE analytical unit of the Roche/Hitachi cobas c systems is intended for the quantitative determination of sodium in serum, plasma or urine using ion-selective electrodes. Sodium measurements are used in the diagnosis and treatment of aldosteronism (excessive secretion of the hormone aldosterone), diabetes insipidus (chronic excretion of large amounts of dilute urine, accompanied by extreme thirst), adrenal hypertension, Addison's disease (caused by destruction of the adrenal glands), dehydration, inappropriate antidiuretic hormone secretion, or other diseases involving electrolyte imbalance. Methadone II (MDN2) is an in vitro diagnostic test for the qualitative and semiquantitative detection of methadone in human urine on Roche/Hitachi cobas c systems at a cutoff concentration of 300 ng/mL. Semiquantitative test results may be obtained that permit laboratories to assess assay performance as part of a quality control program. Semiquantitative assays are intended to determine an appropriate dilution of the specimen for confirmation by a confirmatory method such as gas chromatography/mass spectrometry (GC-MS). Elecsys TSH is an immunoassay for the in vitro quantitative determination of thyrotropin in human serum and plasma. Measurements of TSH are used in the diagnosis of thyroid and pituitary disorders. The electrochemiluminescence immunoassay "ECLIA" is intended for use on cobas e immunoassay analyzers.

## Device Story

cobas pure integrated solutions is a fully automated, software-controlled, random-access clinical analyzer for low-to-mid throughput laboratories. It consolidates clinical chemistry (cobas c 303 unit), ion-selective electrode (ISE) measurements, and immunoassay (cobas e 402 unit) into one system. Inputs include human serum, plasma, urine, or CSF samples. The system uses hexokinase-based spectrophotometry for glucose, ion-selective membranes for sodium, kinetic interaction of microparticles in solution (KIMS) for methadone, and electrochemiluminescence (ECLIA) for TSH. The device processes samples automatically, providing quantitative or qualitative results to healthcare providers. These results assist clinicians in diagnosing metabolic, electrolyte, and endocrine disorders, and in monitoring drug presence. The system benefits patients by enabling rapid, consolidated diagnostic testing in clinical settings.

## Clinical Evidence

Bench testing only. Precision (CLSI EP05-A3), linearity, interference (CLSI EP07), and method comparison (CLSI EP09c) performed. Precision CVs generally <5%. Linearity confirmed across claimed ranges. No significant interference from endogenous substances or common drugs. Method comparison against predicate showed high correlation (r ≥ 0.99).

## Technological Characteristics

Fully automated, random-access, software-controlled analyzer. Analytical units: cobas c 303 (photometric/ISE) and cobas e 402 (ECLIA). Sensing: spectrophotometry (340 nm, 546 nm), ion-selective electrodes, and photomultiplier-based chemiluminescence. Connectivity: networked (cobas link). Software-controlled pipetting, incubation, and detection. Reagents: liquid, ready-to-use.

## Regulatory Identification

A glucose test system is a device intended to measure glucose quantitatively in blood and other body fluids. Glucose measurements are used in the diagnosis and treatment of carbohydrate metabolism disorders including diabetes mellitus, neonatal hypoglycemia, and idiopathic hypoglycemia, and of pancreatic islet cell carcinoma.

## Special Controls

*Classification.* Class II (special controls). The device, when it is solely intended for use as a drink to test glucose tolerance, is exempt from the premarket notification procedures in subpart E of part 807 of this chapter subject to the limitations in § 862.9.

## Predicate Devices

- cobas pro integrated solutions ([K191899](/device/K191899.md))
- Glucose HK Gen.3 ([K191899](/device/K191899.md))
- ISE indirect Na for Gen.2 ([K191899](/device/K191899.md))
- ONLINE DAT Methadone II ([K021505](/device/K021505.md))
- Elecsys TSH ([K191899](/device/K191899.md))

## Submission Summary (Full Text)

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>
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FDA

U.S. FOOD &amp; DRUG

ADMINISTRATION

# 510(k) SUBSTANTIAL EQUIVALENCE DETERMINATION DECISION SUMMARY

ASSAY AND INSTRUMENT

## I Background Information:

A 510(k) Number

K220134

B Applicant

Roche Diagnostics

C Proprietary and Established Names

Glucose HK Gen.3, ISE indirect Na for Gen.2, ONLINE DAT Methadone II, Elecsys TSH, cobas pure integrated solutions

D Regulatory Information

|  Product Code(s) | Classification | Regulation Section | Panel  |
| --- | --- | --- | --- |
|  CFR | Class II | 21 CFR 862.1345 - Glucose Test System | CH - Clinical Chemistry  |
|  JGS | Class II | 21 CFR 862.1665 - Sodium test system | CH - Clinical Chemistry  |
|  JLW | Class II | 21 CFR 862.1690 - Thyroid stimulating hormone test system | CH - Clinical Chemistry  |
|  DJR | Class II | 21 CFR 862.3620 - Methadone test system | TX - Clinical Toxicology  |
|  JJE | Class I | 21 CFR 862.2160 - Discrete photometric chemistry analyzer for clinical use | CH - Clinical Chemistry  |

Food and Drug Administration

10903 New Hampshire Avenue

Silver Spring, MD 20993-0002

www.fda.gov

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K220134 - Page 2 of 34

## II Submission/Device Overview:

### A Purpose for Submission:
Addition of previously cleared assays to a new instrument platform

### B Measurand:
Glucose, Sodium, Methadone, and TSH

### C Type of Test:
Quantitative photometric, ion selective electrode, and immunoassay

## III Intended Use/Indications for Use:

### A Intended Use(s):
See Indications for Use below.

### B Indication(s) for Use:
cobas pure integrated solutions is an automated analyzer, intended for running qualitative, semi-quantitative and quantitative clinical chemistry and immunochemistry assays as well as ion selective measurements.

Glucose HK Gen.3 is an in vitro test for the quantitative determination of glucose in human serum, plasma, urine and CSF on Roche/Hitachi cobas c systems. Glucose measurements are used in the diagnosis and treatment of carbohydrate metabolism disorders including diabetes mellitus, neonatal hypoglycemia, idiopathic hypoglycemia and pancreatic islet cell tumors.

The ISE analytical unit of the Roche/Hitachi cobas c systems is intended for the quantitative determination of sodium in serum, plasma or urine using ion-selective electrodes. Sodium measurements are used in the diagnosis and treatment of aldosteronism (excessive secretion of the hormone aldosterone), diabetes insipidus (chronic excretion of large amounts of dilute urine, accompanied by extreme thirst), adrenal hypertension, Addison's disease (caused by destruction of the adrenal glands), dehydration, inappropriate antidiuretic hormone secretion, or other diseases involving electrolyte imbalance.

Methadone II (MDN2) is an in vitro diagnostic test for the qualitative and semiquantitative detection of methadone in human urine on Roche/Hitachi cobas c systems at a cutoff concentration of 300 ng/mL. Semiquantitative test results may be obtained that permit laboratories to assess assay performance as part of a quality control program. Semiquantitative assays are intended to determine an appropriate dilution of the specimen for confirmation by a confirmatory method such as gas chromatography/mass spectrometry (GC-MS).

Elecsys TSH is an immunoassay for the in vitro quantitative determination of thyrotropin in human serum and plasma. Measurements of TSH are used in the diagnosis of thyroid and

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pituitary disorders. The electrochemiluminescence immunoassay "ECLIA" is intended for use on cobas e immunoassay analyzers.

## C Special Conditions for Use Statement(s):

Rx - For Prescription Use Only

## D Special Instrument Requirements:

cobas pure integrated solutions analyzer

## IV Device/System Characteristics:

## A Device Description:

### cobas pure Integrated Solutions System

The cobas pure integrated solutions is a fully automated, random-access, software-controlled system intended for in vitro quantitative, semi-quantitative and qualitative analysis of analytes in body fluids. The system consolidates clinical chemistry, homogenous and heterogeneous immunoassays as well as electrolyte testing within one workplace. The cobas pure integrated solutions consists of a clinical chemistry analytical unit (cobas c 303) with an integrated ISE analytical unit, an immunoassay analytical unit (cobas e 402) and a core unit.

### Glucose HK Gen. 3

The reagent working solutions include:

- R1: MES buffer: 5.0 mmol/L, pH 6.0; Mg²⁺, 24 mmol/L; ATP, ≥ 4.5 mmol/L; NADP, ≥ 7.0 mmol/L; preservative
- R3: HEPES buffer: 200 mmol/L, pH 8.0; Mg²⁺, 4 mmol/L; HK (yeast), ≥ 300 μkat/L; G-6-PDH (E. coli), ≥ 300 μkat/L; preservative

### ISE indirect Na for Gen. 2

ISE auxiliary reagents include:

- ISE Reference Electrolyte: 1 mol/L potassium chloride
- ISE Diluent: HEPES buffer, 10 mmol/L; triethanolamine, 7 mmol/L; preservative
- ISE Internal Standard: HEPES buffer, 10 mmol/L; triethanolamine, 7 mmol/L; sodium chloride, 3.06 mmol/L; sodium acetate, 1.45 mmol/L; potassium chloride, 0.16 mmol/L; preservative
- ISE Cleaning Solution: sodium hydroxide solution, 3 mol/L with sodium hypochlorite solution &lt; 2 % active Cl
- ISE Deproteinizer: sodium hydroxide solution, approximately 1.2 % active Cl
- Electrodes: Sodium, reference

### ONLINE DAT Methadone II

The reagent working solutions include:

R1: Conjugated methadone derivative; buffer; bovine serum albumin; 0.09 % sodium azide

R2: Microparticles attached to methadone antibody (mouse monoclonal); buffer; bovine serum albumin; 0.09 % sodium azide

K220134 - Page 3 of 34

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K220134 - Page 4 of 34

# Elecsys TSH

The reagent working solutions include:

- M: streptavidin-coated microparticles, 1 bottle, 14.1 mL: streptavidin-coated microparticles 0.72 mg/mL; preservative
- R1: Anti-TSH-Ab~biotin, 1 bottle, 15.8 mL: biotinylated monoclonal anti TSH antibody (mouse) 2.0 mg/L; phosphate buffer 100 mmol/L, pH 7.2; preservative
- R2: Anti-TSH-Ab~Ru(bpy)₂³⁺, 1 bottle, 13.9 mL: monoclonal anti TSH antibody (mouse/human) labeled with ruthenium complex 1.5 mg/L; phosphate buffer 100 mmol/L, pH 7.2; preservative

## B Principle of Operation:

### Glucose HK Gen. 3

Glucose is phosphorylated by hexokinase (HK) in the presence of adenosine triphosphate (ATP) and magnesium ions to produce glucose-6-phosphate (G-6-P) and adenosine diphosphate (ADP). Glucose-6-phosphate dehydrogenase (G-6-PDH) specifically oxidizes G-6-P to 6-phosphogluconate with the concurrent reduction of nicotinamide adenine dinucleotide (NAD) to nicotinamide adenine dinucleotide reduced (NADH). One micromole of NADH is produced for each micromole of glucose consumed. The NADH produced absorbs light at 340 nm and can be detected spectrophotometrically as an increased absorbance. The rate of NADPH formation during the reaction is directly proportional to the glucose concentration.

### ISE indirect Na for Gen. 2

The ion-selective electrode (ISE) analytical unit for Na⁺ employs an ion-selective membrane to develop an electrical potential (electromotive force, EMF) for the measurements of ions in solution. The selective membrane is in contact with both the test solution and an internal filling solution. Due to the selectivity of the membrane, only the ions to be measured contribute to the EMF. The membrane EMF is determined by the difference in concentration of the test ion in the test solution and the internal filling solution. The EMF develops and ion concentration is determined according to the Nernst equation.

### ONLINE DAT Methadone II

The Methadone assay is an immunoassay based on the kinetic interaction of microparticles in a solution (KIMS) as measured by changes in light transmission. In the absence of sample drug, soluble drug conjugates bind to antibody-bound microparticles, causing the formation of particle aggregates. As the aggregation reaction proceeds in the absence of sample drug, the absorbance at 546 nm increases.

When a urine sample contains the drug in question, this drug competes with the drug derivative conjugate for microparticle-bound antibody. Antibody bound to sample drug is no longer available to promote particle aggregation, and subsequent particle lattice formation is inhibited. The presence of sample drug diminishes the increasing absorbance in proportion to the concentration of drug in the sample. Sample drug content is determined relative to the value obtained for a known cutoff concentration of drug.

### Elecsys TSH

The Elecsys TSH immunoassay makes use of a sandwich test principle using monoclonal antibodies specifically directed against human TSH. The antibodies labeled with ruthenium

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complex consist of a chimeric construct from human and mouse specific components. In the first incubation step, a biotinylated monoclonal TSH-specific antibody and a monoclonal TSH-specific antibody labeled with a ruthenium complex react to form a sandwich complex. In the second incubation step, after addition of streptavidin-coated microparticles, the complex becomes bound to the solid phase via interaction of biotin and streptavidin. The reaction mixture is then aspirated into the measuring cell where the microparticles are magnetically captured on the surface of the electrode. Unbound substances are removed by washing with ProCell II M. Application of a voltage to the electrode then induces chemiluminescent emission which is measured and amplified by a photomultiplier. Results are determined via a calibration curve which is instrument specific, generated by 2-point calibration and a master curve provided via the cobas link.

## C Instrument Description Information:

1. Instrument Name:
cobas pure integrated solutions

2. Specimen Identification:
The specimen is in a tube with a barcode label. The system identifies the specimen by scanning the barcode. This is the primary use case for sample identification. The system also supports manual entry of sample IDs to support the needs of the customer.

3. Specimen Sampling and Handling:
Specimen sampling and handling procedures are analyte specific and documented in the respective reagent method sheets.

4. Calibration:
The software of the cobas pure integrated solution automatically recommends calibration for all tests requiring calibration. Calibration methods and procedures are analyte specific and documented in the respective reagent method sheets.

5. Quality Control:
Quality control procedures are analyte specific and documented in the respective reagent method sheets.

## V Substantial Equivalence Information:

### A Predicate Device Name(s):

Glucose HK Gen.3, ISE indirect Na for Gen.2, Elecsys TSH, ONLINE DAT Methadone II, cobas pro integrated solutions

K220134 - Page 5 of 34

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B Predicate 510(k) Number(s):

K191899, K021505

C Comparison with Predicate(s):

Comparison of the Glucose HK Gen. 3

|  Device & Predicate Device(s): | K220134 | K191899  |
| --- | --- | --- |
|  Device Trade Name | Glucose HK Gen. 3 | Glucose HK Gen. 3  |
|  General Device Characteristic Similarities |  |   |
|  Intended Use/Indications for Use | In vitro test for the quantitative determination of glucose in human serum, plasma, urine, and CSF on Roche/Hitachi cobas c systems | Same  |
|  Technology | Photometric | Same  |
|  Test Format | Enzymatic | Same  |
|  Test Type | Quantitative | Same  |
|  Assay Protocol | R1+R2+Diluent+Sample, incubation | Same  |
|  Sample Volume | 1.5 μL | Same  |
|  Handling of R1 and R2 | Liquid, ready to use | Same  |
|  Measuring Range | 2.0-750 mg/dL (0.11-41.6 mmol/L) | Same  |
|  General Device Characteristic Differences |  |   |
|  Instrument Platform | cobas c 303 in cobas pure integrated solutions | cobas c 503 in cobas pro integrated solutions  |

Comparison of the ISE indirect Na for Gen.2

|  Device & Predicate Device(s): | K220134 | K191899  |
| --- | --- | --- |
|  Device Trade Name | ISE indirect Na for Gen.2 | ISE indirect Na for Gen.2  |
|  General Device Characteristic Similarities |  |   |
|  Intended Use/Indications for Use | In vitro test for the quantitative determination of sodium in serum, plasma or urine | Same  |

K220134 - Page 6 of 34

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Comparison of the ONLINE DAT Methadone II

|  Device & Predicate Device(s): | K220134 | K021505  |
| --- | --- | --- |
|  Device Trade Name | ONLINE DAT Methadone II | ONLINE DAT Methadone II  |
|  General Device Characteristic Similarities |  |   |
|  Intended Use/Indications for Use | In vitro test for the qualitative and semiquantitative detection of methadone in human urine on Roche/Hitachi cobas c systems at a cutoff concentration of 300 ng/mL | Same  |
|  Technology | KIMS, Kinetic interaction of microparticles in a solution | Same  |
|  Reagents | Conjugate Working Solution: Conjugated methadone derivative in buffer with BSA and preservative. Antibody/Microparticle Working Solution: | Same  |

K220134 - Page 7 of 34

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Comparison of the Elecsys TSH

|  Device & Predicate Device(s): | K220134 | K021505  |
| --- | --- | --- |
|   | Microparticles coated with methadone monoclonal antibody (mouse) in buffer with BSA and preservative. |   |
|  Qualitative and Semi-Quantitative Cutoffs | 300 ng/mL | Same  |
|  General Device Characteristic Differences |  |   |
|  Instrument Platform | cobas c 303 in cobas pure integrated solutions | cobas c 503 in cobas pro integrated solutions  |
|  Device & Predicate Device(s): | K220134 | K191899  |
| --- | --- | --- |
|  Device Trade Name | Elecsys TSH | Elecsys TSH  |
|  General Device Characteristic Similarities |  |   |
|  Intended Use/Indications for Use | Immunoassay for the in vitro quantitative determination of thyrotropin in human serum and plasma | Same  |
|  Technology | Electrochemiluminescence Immunoassay (ECLIA) | Same  |
|  Test Format | Sandwich | Same  |
|  Test type | Quantitative | Same  |
|  Antibody/Reagents | Biotinylated monoclonal anti-TSH antibody (mouse) Monoclonal anti-TSH antibody (mouse/human) labeled with ruthenium complex Streptavidin –coated microparticles | Same  |
|  Assay Protocol | R1+R2+sample, incubation, + beads, incubation | Same  |
|  Sample Volume | 50 μL | Same  |
|  Handling of R1 and R2 | Liquid, ready to use | Same  |
|  Measuring Range | 0.005-100 μIU/mL | Same  |
|  Biotin Tolerance | 1200 ng/mL | Same  |

K220134 - Page 8 of 34

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Comparison of the cobas pure integrated solutions to the cobas pro integrated solutions systems

|  Device & Predicate Device(s): | K220134 | K191899  |
| --- | --- | --- |
|  Device Trade Name | cobas pure integrated solutions | cobas pro integrated solutions  |
|  General Device Characteristic Similarities |  |   |
|  Intended Use/Indications for Use | Automated analyzer, intended for running qualitative, semi-quantitative and quantitative clinical chemistry and immunochemistry assays as well as ion selective measurements | Same  |
|  Function Performed | Data Input, Sample Processing, Result Calculation, Result Reporting, Quality Control, Infrastructure (power, water supply) | Same  |
|  PC (Controller Unit) Functions | Data Input (Touch screen, Disc), Data Output (Screen, printer) | Same  |
|  Core Unit Functions | Real time database, data input and output (via HOST communication), control of sample conveyer | Same  |
|  Analytical Unit(s) Functions | Control of analytic processes (pipetting, incubation, detection) Primary signal processing | Same  |
|  Data Storage | Real time database in Core Unit (storage of System and Application | Same  |

K220134 - Page 9 of 34

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|  Device & Predicate Device(s): | K220134 | K191899  |
| --- | --- | --- |
|   | parameters, Calibration Data, QC Data, Sample Results, Alarm history) |   |
|  Result Calculation | Automated measuring of signal using various methods according to automated calculation of concentration via calibration curve | Same  |
|  Initial Cassette Volume Check (ICVC) for Reagent Pipetting | Available | Same  |
|  Data Concept (Application Parameter, Calibrator, Control Value Transfer) | Electronic transfer possible (user must accept transfer before parameter applied) | Same  |
|  General Device Characteristic Differences |  |   |
|  Software | cobas pure integrated solutions software | cobas pro integrated solutions software  |
|  Configuration | Up to 2 analytical units with one PC and one sample supply unit | Several analytical units with one PC and one Core unit  |
|  Units Controlled | cobas c 303 analytical unit with integrated ISE analytical unit, cobas e 402 | cobas c 503, cobas e 801, and cobas pro ISE analyzers  |
|  Throughput | cobas c 303: 450 tests/hr
cobas c 303 ISE: 450 tests/hr
cobas e 402: 120 tests/hr | cobas c 503: 1000 tests/hr
cobas pro ISE: 900 tests/hr
cobas e 801: 300 tests/hr  |

VI Standards/Guidance Documents Referenced:

Clinical and Laboratory Standards Institute (CLSI) EP05-A3 Evaluation of Precision of Quantitative Measurement Procedures; Approved Guideline - Third Edition

CLSI EP07 Interference Testing in Clinical Chemistry – Third Edition

CLSI EP09c Measurement Procedure Comparison and Bias Estimation Using Patient Samples – Third Edition

CLSI EP17-A2 Evaluation of Detection Capability for Clinical Laboratory Measurement Procedures; Approved Guideline - Second Edition

K220134 - Page 10 of 34

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VII Performance Characteristics (if/when applicable):

# A Analytical Performance:

# 1. Precision/Reproducibility:

Repeatability (within-run precision) and intermediate precision (within-laboratory precision) were evaluated following the recommendations in the CLSI guideline EP05-A3.

# Glucose HK Gen.3: Repeatability and Intermediate Precision

The precision of the Glucose HK Gen.3 assay was evaluated on one cobas pure integrated solutions system using three lots of reagent. The protocol consisted of testing two sample replicates per run, two runs per day for 21 days. Repeatability and within-lab precision were calculated. The samples were randomized in each run separately. Two serum-based controls and five levels of individual serum samples were used (Serum 1 was diluted, Serum 2 was native, and Serum 3-5 were spiked). The protocol was repeated for each application (serum, urine, CSF). For urine, individual samples were also used (Urine 1 was native and Urine 2-5 were spiked) while for CSF, pooled samples were used (CSF 1-2 were diluted, CSF 3 was native, and CSF 4-5 were spiked). The results for a representative reagent lot are summarized in the table below.

|  Sample | N | Mean (mg/dL) | Repeatability |   | Mean (mg/dL) | Within-Laboratory Precision  |   |
| --- | --- | --- | --- | --- | --- | --- | --- |
|   |   |   |  SD (mg/dL) | CV (%) |   | SD (mg/dL) | CV (%)  |
|  Serum Control 1 | 84 | 100 | 0.292 | 0.3 | 100 | 0.611 | 0.6  |
|  Serum Control 2 | 84 | 238 | 0.901 | 0.4 | 238 | 1.33 | 0.6  |
|  Serum 1 | 84 | 3.73 | 0.082 | 2.2 | 3.77 | 0.106 | 2.8  |
|  Serum 2 | 84 | 64.5 | 0.338 | 0.5 | 64.5 | 0.389 | 0.6  |
|  Serum 3 | 84 | 108 | 0.333 | 0.3 | 108 | 0.517 | 0.5  |
|  Serum 4 | 84 | 384 | 1.21 | 0.3 | 389 | 1.63 | 0.4  |
|  Serum 5 | 84 | 715 | 2.83 | 0.4 | 724 | 3.46 | 0.5  |
|  Urine Control 1 | 84 | 29.2 | 0.214 | 0.7 | 29.2 | 0.241 | 0.8  |
|  Urine Control 2 | 84 | 294 | 1.46 | 0.5 | 292 | 1.55 | 0.5  |
|  Urine 1 | 84 | 3.12 | 0.154 | 5.0 | 3.12 | 0.163 | 5.2  |
|  Urine 2 | 84 | 13.1 | 0.138 | 1.1 | 13.2 | 0.168 | 1.3  |
|  Urine 3 | 84 | 70.8 | 0.290 | 0.4 | 70.8 | 0.323 | 0.5  |
|  Urine 4 | 84 | 377 | 1.35 | 0.4 | 377 | 1.67 | 0.4  |
|  Urine 5 | 84 | 732 | 2.83 | 0.4 | 732 | 3.17 | 0.4  |
|  CSF Control 1 | 84 | 62.5 | 0.333 | 0.5 | 62.5 | 0.564 | 0.9  |
|  CSF Control 2 | 84 | 31.9 | 0.238 | 0.7 | 31.9 | 0.301 | 0.9  |

K220134 - Page 11 of 34

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|  Sample | N | Mean (mg/dL) | Repeatability |   | Mean (mg/dL) | Within-Laboratory Precision  |   |
| --- | --- | --- | --- | --- | --- | --- | --- |
|   |   |   |  SD (mg/dL) | CV (%) |   | SD (mg/dL) | CV (%)  |
|  CSF 1 | 84 | 3.71 | 0.108 | 2.9 | 3.71 | 0.110 | 3.0  |
|  CSF 2 | 84 | 38.2 | 0.234 | 0.6 | 38.2 | 0.333 | 0.9  |
|  CSF 3 | 84 | 72.3 | 0.441 | 0.6 | 72.3 | 0.627 | 0.9  |
|  CSF 4 | 84 | 374 | 2.38 | 0.6 | 375 | 3.14 | 0.8  |
|  CSF 5 | 84 | 728 | 4.16 | 0.6 | 728 | 5.33 | 0.7  |

# ISE Indirect Na for Gen. 2: Repeatability and Intermediate Precision

The precision of the ISE indirect Na for Gen. 2 test was evaluated on one cobas pure integrated solutions system using one reagent lot. The protocol consisted of testing two sample replicates per run, two runs per day for 21 days. Repeatability and within-lab precision were calculated. The samples were randomized in each run separately. The protocol was repeated for each application (serum, plasma and urine). Pooled samples were used for each level, were aliquoted and then measured. Samples 1 and 5 were diluted or spiked, samples 2-4 were native. The results are summarized in the table below.

|  Sample | N | Mean (mmol/L) | Repeatability |   | Within-Laboratory Precision  |   |
| --- | --- | --- | --- | --- | --- | --- |
|   |   |   |  SD (mmol/L) | CV (%) | SD (mmol/L) | CV (%)  |
|  Serum Control 1 | 84 | 112 | 0.665 | 0.6 | 1.13 | 1.0  |
|  Serum Control 2 | 84 | 137 | 0.683 | 0.5 | 1.20 | 0.9  |
|  Serum 1 | 84 | 86.1 | 0.458 | 0.5 | 1.18 | 1.4  |
|  Serum 2 | 84 | 132 | 0.561 | 0.4 | 1.06 | 0.8  |
|  Serum 3 | 84 | 136 | 0.688 | 0.5 | 0.95 | 0.7  |
|  Serum 4 | 84 | 157 | 0.834 | 0.5 | 1.13 | 0.7  |
|  Serum 5 | 84 | 174 | 0.832 | 0.5 | 1.31 | 0.8  |
|  Plasma Control 1 | 84 | 112 | 0.493 | 0.4 | 1.07 | 1.0  |
|  Plasma Control 2 | 84 | 137 | 0.525 | 0.4 | 1.16 | 0.9  |
|  Plasma 1 | 84 | 87.4 | 0.411 | 0.5 | 1.09 | 1.2  |
|  Plasma 2 | 84 | 131 | 0.584 | 0.4 | 0.98 | 0.8  |
|  Plasma 3 | 84 | 136 | 0.597 | 0.4 | 1.07 | 0.8  |
|  Plasma 4 | 84 | 156 | 0.721 | 0.5 | 0.96 | 0.6  |
|  Plasma 5 | 84 | 172 | 0.662 | 0.4 | 1.16 | 0.7  |
|  Urine Control 1 | 84 | 82.3 | 0.394 | 0.5 | 0.95 | 1.2  |
|  Urine Control 2 | 84 | 178 | 0.823 | 0.5 | 1.15 | 0.6  |
|  Urine 1 | 84 | 26.8 | 0.267 | 1.0 | 1.16 | 4.3  |
|  Urine 2 | 84 | 136 | 0.557 | 0.4 | 0.79 | 0.6  |

K220134 - Page 12 of 34

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|  Sample | N | Mean (mmol/L) | Repeatability |   | Within-Laboratory Precision  |   |
| --- | --- | --- | --- | --- | --- | --- |
|   |   |   |  SD (mmol/L) | CV (%) | SD (mmol/L) | CV (%)  |
|  Urine 3 | 84 | 112 | 0.433 | 0.4 | 0.81 | 0.7  |
|  Urine 4 | 84 | 199 | 0.942 | 0.5 | 1.69 | 0.8  |
|  Urine 5 | 84 | 239 | 0.868 | 0.4 | 2.50 | 1.0  |

# ONLINE DAT Methadone II

The precision of the ONLINE DAT Methadone II test was evaluated on one cobas pure integrated solutions system using three reagent lots using both the qualitative and the semi-quantitative mode. The protocol consisted of testing two sample replicates per run, two runs per day for 21 days. Seven human samples and four control samples were used. The samples with the concentrations provided in the tables below were prepared by spiking drug into negative urine. The analyte concentration was confirmed using a validated method. The results for a representative reagent lot are summarized in the tables below.

Summary of Within-Lab Precision - Qualitative (300 ng/mL cutoff):

|  Sample | N | Negative | Positive  |
| --- | --- | --- | --- |
|  Sample -100% (zero) | 84 | 84 | 0  |
|  Sample -75% | 84 | 84 | 0  |
|  Sample -50% | 84 | 84 | 0  |
|  Control 1 | 84 | 84 | 0  |
|  Control 2 | 84 | 84 | 0  |
|  Cutoff | 84 | 9 | 75  |
|  Control 3 | 84 | 0 | 84  |
|  Control 4 | 84 | 0 | 84  |
|  Sample +50% | 84 | 0 | 84  |
|  Sample +75% | 84 | 0 | 84  |
|  Sample +100% | 84 | 0 | 84  |

Summary of Precision Results - Semi-Quantitative (300 ng/mL cutoff):

|  Sample | N | Mean (mmol/L) | Repeatability |   | Within-Laboratory Precision  |   |
| --- | --- | --- | --- | --- | --- | --- |
|   |   |   |  SD (ng/mL) | CV (%) | SD (ng/mL) | CV (%)  |
|  Sample - 100% (zero) | 84 | 22.6 | 11.6 | 51.5 | 17.7 | 78.3  |
|  Sample - 75% | 84 | 112 | 6.70 | 6.0 | 10.3 | 9.1  |
|  Sample - 50% | 84 | 155 | 5.31 | 3.4 | 9.81 | 6.3  |
|  Control 1 | 84 | 234 | 6.78 | 2.9 | 11.0 | 4.7  |
|  Control 2 | 84 | 236 | 5.68 | 2.4 | 10.5 | 4.5  |
|  Cutoff | 84 | 314 | 6.19 | 2.0 | 13.5 | 4.3  |
|  Control 3 | 84 | 389 | 7.00 | 1.8 | 15.3 | 3.9  |
|  Control 4 | 84 | 379 | 7.72 | 2.0 | 14.9 | 3.9  |

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|  Sample | N | Mean (mmol/L) | Repeatability |   | Within-Laboratory Precision  |   |
| --- | --- | --- | --- | --- | --- | --- |
|   |   |   |  SD (ng/mL) | CV (%) | SD (ng/mL) | CV (%)  |
|  Sample +50% | 84 | 484 | 9.21 | 1.9 | 19.5 | 4.0  |
|  Sample +75% | 84 | 543 | 10.7 | 2.0 | 20.5 | 3.8  |
|  Sample +100% | 84 | 632 | 12.3 | 1.9 | 21.9 | 3.5  |

## Elecsys TSH

Precision of the Elecsys TSH assay was evaluated on one cobas pure integrated solutions system using one reagent lot. The protocol consisted of testing two sample replicates per run, two runs per day for 21 days. Repeatability and within-lab precision were calculated. The samples were run in randomized order on the analyzer. Three serum-based controls and five levels of pooled serum samples were used (Serum 1-4 were native, Serum 5 was spiked with recombinant TSH). The results are summarized in the table below.

|  Sample | N | Mean (μIU/mL) | Repeatability |   | Within-Laboratory Precision  |   |
| --- | --- | --- | --- | --- | --- | --- |
|   |   |   |  SD (μIU/mL) | CV (%) | SD (μIU/mL) | CV (%)  |
|  Serum Control 1 | 84 | 1.30 | 0.013 | 1.0 | 0.016 | 1.2  |
|  Serum Control 2 | 84 | 7.89 | 0.069 | 0.9 | 0.111 | 1.4  |
|  Serum Control 3 | 84 | 0.169 | 0.002 | 0.9 | 0.002 | 1.3  |
|  Serum 1 | 84 | 0.012 | 0.001 | 4.5 | 0.001 | 5.2  |
|  Serum 2 | 84 | 0.267 | 0.003 | 1.0 | 0.004 | 1.6  |
|  Serum 3 | 82 | 3.77 | 0.046 | 1.2 | 0.055 | 1.5  |
|  Serum 4 | 84 | 55.5 | 0.378 | 0.7 | 0.601 | 1.1  |
|  Serum 5 | 84 | 97.5 | 0.566 | 0.6 | 0.731 | 0.7  |

## 2. Linearity:

## Glucose HK Gen.3

Linearity of the Glucose HK Gen.3 assay was assessed on one cobas pure integrated solutions system in one run using three reagent lots, measuring three replicates per sample. Four high analyte concentration human serum, plasma (collected in  $\mathrm{K}_2\mathrm{EDTA}$  tubes), urine and CSF samples were diluted to obtain 12-14 levels spanning the measuring range. The results of the linear regression analysis for a representative reagent lot are summarized in the table below.

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|  Sample | Tested Range (mg/dL) | Linear Regression Data | Claimed Linear Range (mg/dL)  |
| --- | --- | --- | --- |
|  Serum | 0-794.7 | y = 1.003x - 0.0273, R² = 0.9998 | 2-750  |
|  Plasma | 0-807.3 | y = 1.000x - 0.0011, R² = 0.9999 | 2-750  |
|  Urine | 0-827.1 | y = 1.002x - 0.0049, R² = 0.9999 | 2-750  |
|  CSF | 0-787.5 | y = 1.001x - 0.0042, R² = 0.9998 | 2-750  |

The results demonstrated linearity of the claimed measuring range for serum, plasma, urine, and CSF (2.0 to  $750\mathrm{mg / dL}$  or 0.11 to  $41.6\mathrm{mmol / L}$ ).

# ISE Indirect Na for Gen. 2

Linearity of the ISE Indirect Na for Gen.2 test was assessed on one cobas pure integrated solutions system in one run using one reagent lot, measuring three replicates per sample. Three high analyte concentration human serum, plasma (collected in Li-heparin tubes) and urine samples were diluted to obtain 11 levels spanning the measuring range. The results of the linear regression analysis are summarized in the table below.

|  Sample | Tested Range (mmol/L) | Linear Regression | Claimed Linear Range (mmol/L)  |
| --- | --- | --- | --- |
|  Serum | 75.1-190 | y = 1.000x + 0.0000, R² = 0.9996 | 80-180  |
|  Plasma | 73.3-183 | y = 1.000x + 0.0000, R² = 0.9996 | 80-180  |
|  Urine | 19.9-259 | y = 1.000x - 0.0312, R² = 0.9995 | 20-250  |

The results demonstrated linearity of the claimed measuring range (80 mmol/L to 180 mmol/L for serum/plasma, and 25 mmol/L to 250 mmol/L for urine).

# ONLINE DAT Methadone II

Linearity of the semi-quantitative ONLINE DAT Methadone II test was assessed on one cobas pure integrated solutions system in one run using one reagent lot, measuring three replicates per sample. A dilution series was prepared from human urine sample pools (spiked with methadone and diluted with human urine) to obtain 12 levels that span the range of the used calibrators. The results of the linearity/recovery study are summarized in the table below.

|  Expected Value (ng/mL) | Observed Value (ng/mL) | Absolute Deviation (ng/mL) | % Recovery  |
| --- | --- | --- | --- |
|  0.0 | 0.0 | 0.0 |   |
|  39.3 | 3.09 | -36.2 |   |
|  55.5 | 20.1 | -35.4 |   |
|  231 | 209 | -22.0 |   |
|  463 | 447 |  | 96.5  |
|  694 | 714 |  | 102.9  |
|  925 | 1073 |  | 116.0  |
|  1156 | 1181 |  | 102.2  |
|  1388 | 1469 |  | 105.8  |

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|  Expected Value (ng/mL) | Observed Value (ng/mL) | Absolute Deviation (ng/mL) | % Recovery  |
| --- | --- | --- | --- |
|  1619 | 1621 |  | 100.1  |
|  1850 | 1880 |  | 101.6  |
|  2081 | 2167 |  | 104.1  |
|  2313 | 2091 |  | 90.4  |

## Elecsys TSH

Linearity of the Elecsys TSH assay was assessed on one cobas pure integrated solutions system in one run using one reagent lot, measuring three replicates per sample. Three high analyte concentration human serum samples were diluted to obtain 12 levels spanning the measuring range. The results of the weighted linear regression analysis for a representative sample set are summarized in the table below.

|  Sample | Tested Range (μIU/mL) | Linear Regression | Claimed Linear Range (μIU/mL)  |
| --- | --- | --- | --- |
|  Serum | 0.000-109 | y = 1.025x - 0.0013, R² = 0.9859 | 0.004-102  |

The results demonstrated linearity of the claimed measuring range (0.004 - 102 μIU/mL).

## Dilution Recovery Studies

### Glucose HK Gen. 3

A dilution study was performed by diluting three high concentration samples each (serum, urine and CSF) with diluent (NaCl 9%) in a 1:2 ratio. The data support the 1:2 dilution claim in the Method Sheet.

### ISE indirect Na for Gen. 2

A dilution study was performed by diluting three high concentration urine samples with the ISE diluent in a 1:46 ratio. The data support the 1:46 dilution claim in the Method Sheet.

### ONLINE DAT Methadone II

No dilution claims are included in the Method Sheet.

### Elecsys TSH

A dilution study was performed by diluting three high concentration serum samples with the Diluent MultiAssay in a 1:10 ratio. The data support the 1:10 dilution claim in the Method Sheet.

## High Dose Hook Effect

### Elecsys TSH

The high-dose hook effect of the Elecsys TSH assay was assessed on the cobas pure integrated solutions system. Three human serum samples were spiked with analyte (recombinant human TSH) to achieve high TSH concentrations. For each sample, a dilution series was performed. The hook concentration reported corresponds to the highest analyte

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concentration that generates a signal $\geq 10\%$ above the upper limit of the measuring range. The sponsor determined that there is no hook effect up to $1000~\mu \mathrm{IU / mL}$ TSH.

3. Analytical Specificity/Interference:

## Endogenous Interference

The purpose of this study was to evaluate endogenous substances for potential interference when using the Glucose HK Gen.3 assay, the ISE indirect Na for Gen.2 test, the ONLINE DAT Methadone II test and the Elecsys TSH assay on the cobas pure integrated solutions system.

## Glucose HK Gen. 3

The effect on the quantitation of the analyte in the presence of potentially interfering endogenous substances using the Glucose HK Gen.3 assay was determined on the cobas pure integrated solutions system using serum, urine, and CSF samples. Glucose levels of approximately $40\mathrm{mg / dL}$ and $220\mathrm{mg / dL}$ were tested. Significant interference was defined as follows:

- Serum/Plasma: bias between the samples with and without interferent is $&gt; \pm 7\mathrm{mg / dL}$ at glucose concentrations $\leq 70\mathrm{mg / dL}$ and $&gt; \pm 10\%$ at glucose concentrations $&gt;70\mathrm{mg / dL}$
- Urine: bias between the samples with and without interferent is $&gt; \pm 2\mathrm{mg / dL}$ at glucose concentrations $\leq 19.8\mathrm{mg / dL}$ and $&gt; \pm 10\%$ at glucose concentrations $&gt;19.8\mathrm{mg / dL}$
- CSF: bias between the samples with and without interferent is $&gt; \pm 4\mathrm{mg / dL}$ at glucose concentrations $\leq 39.6\mathrm{mg / dL}$ and $&gt; \pm 10\%$ at glucose concentrations $&gt;39.6\mathrm{mg / dL}$

The summary of results is presented in the table below.

|  Potential Interferent |   | Highest Concentration Tested Without Significant Interference  |
| --- | --- | --- |
|  Serum | Albumin | 74.5 g/L  |
|   |  Bilirubin | 62 mg/dL  |
|   |  Ditaurobilirubin | 65 mg/dL  |
|   |  Hemolysis | 1090 mg/dL  |
|   |  IgG | 78.6 g/L  |
|   |  Intralipid | 1126 mg/dL  |
|  Urine | Albumin | 2.50 g/L  |
|   |  Calcium | 12.0 mmol/L  |
|   |  Citrate | 11.0 mmol/L  |
|   |  Creatinine | 88.4 mmol/L  |
|   |  Hemolysis | 788 mg/dL  |
|   |  IgG | 1.10 g/L  |
|   |  Magnesium | 25.0 mmol/L  |
|   |  Oxalate | 1.50 mmol/L  |
|   |  Phosphate | 130 mmol/L  |
|   |  Urea | 1800 mmol/L  |
|   |  Uric Acid | 6.00 mmol/L  |
|   |  Urobilinogen | 0.250 mmol/L  |
|  CSF | Ditaurobilirubin | 81 mg/dL  |

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|  Potential Interferent |   | Highest Concentration Tested Without Significant Interference  |
| --- | --- | --- |
|   | Hemolysis | 1086 mg/dL  |

The sponsor provided information to support that the labeling interference limitations are unchanged from the predicate device (K191899).

## ISE indirect Na for Gen. 2

The effect on the quantitation of the analyte in the presence of potentially interfering endogenous substances using the ISE indirect Na for Gen.2 test was determined on the cobas pure integrated solutions system using human plasma and serum samples. Sodium levels of approximately 124 mmol/L and 151 mmol/L were tested. Significant interference was defined as bias between the samples with and without interferent &gt; ± 10%. The summary of results is presented in the table below.

|  Potential Interferent |   | Highest Concentration Tested Without Significant Interference  |
| --- | --- | --- |
|  Plasma | Bilirubin | 61 mg/dL  |
|   |  Ditaurobilirubin | 61 mg/dL  |
|   |  Hemolysis | 1110 mg/dL  |
|   |  Intralipid | 2043 mg/dL  |
|  Serum | Bilirubin | 60.0 mg/dL  |
|   |  Ditaurobilirubin | 61.0 mg/dL  |
|   |  Hemolysis | 1008 mg/dL  |
|   |  Intralipid | 2017 mg/dL  |

The sponsor provided information to support that the labeling interference limitations are unchanged from the predicate device (K191899).

## ONLINE DAT Methadone II

The effects of potentially interfering endogenous substances on the ONLINE DAT Methadone II test were determined on the cobas pure integrated solutions system using urine samples spiked with methadone. Methadone levels of approximately 225 ng/mL and 375 ng/mL (corresponding to ±25% of the cutoff concentration) were tested. Significant interference was defined as crossover from the expected result for each control level. The summary of results is presented in the table below.

|  Potential Interferent | Concentration tested | Spiked Methadone Concentration  |   |
| --- | --- | --- | --- |
|   |   |  225 ng/mL (Negative Control -25% Cutoff,) | 375 ng/mL (Positive Control, +25% Cutoff)  |
|  Acetone | 800 mg/dL | Neg | Pos  |
|  Albumin | 500 mg/dL | Neg | Pos  |
|  Ascorbic acid | 1500 mg/dL | Neg | Pos  |
|  Bilirubin | 25 mg/dL | Neg | Pos  |
|  Calcium | 48 mg/dL | Neg | Pos  |

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|  Potential Interferent | Concentration tested | Spiked Methadone Concentration  |   |
| --- | --- | --- | --- |
|   |   |  225 ng/mL (Negative Control -25% Cutoff,) | 375 ng/mL (Positive Control, +25% Cutoff)  |
|  Creatinine | 500 mg/dL | Neg | Pos  |
|  Ethanol | 800 mg/dL | Neg | Pos  |
|  Glucose | 2000 mg/dL | Neg | Pos  |
|  Hemoglobin | 750 mg/dL | Neg | Pos  |
|  IgG | 110 mg/dL | Neg | Pos  |
|  Magnesium (MgCl2) | 238 mg/dL | Neg | Pos  |
|  NaCl | 2900 mg/dL and 5800 mg/dL | Neg | Pos  |
|  Oxalic acid | 200 mg/dL | Neg | Pos  |
|  Phosphate (NaH2PO4, Dihydrate) | 2028 mg/dL | Neg | Pos  |
|  Urea | 6000 mg/dL | Neg | Pos  |
|  Uric Acid | 101 mg/dL | Neg | Pos  |
|  Urobilinogen | 15 mg/dL | Neg | Pos  |
|  Citrate | 284 mg/dL | Neg | Pos  |
|  pH | 4-8 | Neg | Pos  |

No interference/crossover was observed from endogenous substances.

## Elecsys TSH

The effect on the quantitation of the analyte in the presence of endogenous interfering substances using the Elecsys TSH assay was determined on the cobas pure integrated solutions system using human serum samples with a low (approximately 0.5 µIU/mL), a mid (approximately 4 µIU/mL) and a high (approximately 7 µIU/mL) TSH concentration. Significant interference was defined as bias between the samples with and without interferent &gt; ± 10%. The summary of results is presented in the table below.

|  Potential Interferent | Highest Concentration Tested Without Significant Interference  |
| --- | --- |
|  Intralipid | 2000 mg/dL  |
|  Biotin | 1200 mg/dL  |
|  Bilirubin | 66 mg/dL  |
|  Hemoglobin | 1000 mg/dL  |
|  Rheumatoid Factor (RF) | 1500 IU/mL  |
|  Human IgG | 3.37 g/dL  |
|  Human IgM | 0.64 g/dL  |

The sponsor provided information to support that the labeling interference limitations are unchanged from the predicate device (K191899).

The following statement regarding biotin interference is included in the labeling: "This assay has no biotin interference in serum concentrations up to 1200 ng/mL. Some studies have shown that serum concentrations of biotin can reach up to 355 ng/mL within the

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first hour after biotin ingestion for subjects consuming supplements of 20 mg biotin per day and up to 1160 ng/mL for subjects after a single dose of 300 mg biotin.

References:
7. Grimsey P, Frey N, Bendig G, et al. Population pharmacokinetics of exogenous biotin and the relationship between biotin serum levels and in vitro immunoassay interference. Int J Pharmacokinet 2017;2(4):247-256.
8. Piketty ML, Prie D, Sedel F, et al. High-dose biotin therapy leading to false biochemical endocrine profiles: validation of a simple method to overcome biotin interference. Clin Chem Lab Med 2017;55(6):817-825.

Exogenous Interference

The purpose of these studies was to evaluate drugs for potential interference when using the Glucose HK Gen.3 assay, the ISE indirect Na for Gen.2 test, the ONLINE DAT Methadone II test and the Elecsys TSH assay on the cobas pure integrated solutions system. The acceptance criteria were recovery within 100 ± 10% for all evaluated assays except methadone where significant interference was defined as crossover from the expected result for each control level.

Glucose HK Gen. 3

The effect on the quantitation of the analyte in the presence of potentially interfering drugs using the Glucose HK Gen.3 assay was determined on the cobas pure integrated solutions system using serum, urine and CSF samples. Glucose levels of approximately 40 mg/dL and 220 mg/dL were tested. The summary of results is presented in the table below.

|  Potential Interferent |   | Highest Concentration Tested Without Significant Interference  |
| --- | --- | --- |
|  Serum | N-Acetylcysteine | 150 mg/L  |
|   |  Acetylsalicylic acid | 30 mg/L  |
|   |  Ampicillin-Na | 75 mg/L  |
|   |  Ascorbic acid | 52.5 mg/L  |
|   |  Cefoxitin | 750 mg/L  |
|   |  Doxicyclinc | 18.0 mg/L  |
|   |  Heparin | 3300 IU/L  |
|   |  Levodopa | 7.5 mg/L  |
|   |  Methyldopa | 22.5 mg/L  |
|   |  Metronidazole | 123 mg/L  |
|   |  Rifampicin | 48 mg/L  |
|   |  Acetaminophen | 156 mg/L  |
|   |  Cyclosporine | 1.8 mg/L  |
|   |  Ibuprofen | 219 mg/L  |
|   |  Theophylline | 60 mg/L  |
|   |  Phenylbutazone | 321 mg/L  |
|  Urine | Acetaminophen | 3000 mg/L  |
|   |  N-Acetylcysteine | 10 mg/L  |
|   |  Ascorbic acid | 4000 mg/L  |

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|  Potential Interferent |   | Highest Concentration Tested Without Significant Interference  |
| --- | --- | --- |
|   | Cefoxitin | 12000 mg/L  |
|   |  Gentamycine sulfate | 400 mg/L  |
|   |  Levodopa | 1000 mg/L  |
|   |  Methyldopa | 2000 mg/L  |
|   |  Ofloxacine | 900 mg/L  |
|   |  Ibuprofen | 4000 mg/L  |
|   |  Phenazopyridine | 300 mg/L  |
|   |  Salicyluric acid | 100 mg/L  |
|   |  Tetracycline | 100 mg/L  |

## ISE indirect Na for Gen. 2

The effect on the quantitation of the analyte in the presence of potentially interfering drugs using the ISE indirect Na for Gen. 2 assay was determined on the cobas pure integrated solutions system using human plasma, serum, and urine samples. Sodium levels of approximately 124 and 156 mmol/L for serum and plasma and at 32 and 197 mmol/L for urine were tested. The summary of results is presented in the table below.

|  Potential Interferent |   | Highest Concentration Tested Without Significant Interference  |
| --- | --- | --- |
|  Serum and Plasma | N-Acetylcysteine | 1660 mg/L  |
|   |  Ampicillin-Na | 1000 mg/L  |
|   |  Ascorbic acid | 300 mg/L  |
|   |  Cyclosporine | 5 mg/L  |
|   |  Na-Cefoxitin | 2500 mg/L  |
|   |  Heparin | 5000 IU/L  |
|   |  Intralipid | 10000 mg/L  |
|   |  Levodopa | 20 mg/L  |
|   |  Methyldopa | 20 mg/L  |
|   |  Metronidazole | 200 mg/L  |
|   |  Phenylbutazone | 400 mg/L  |
|   |  Doxycycline | 50 mg/L  |
|   |  Acetylsalicylic acid | 1000 mg/L  |
|   |  Rifampicin | 60 mg/L  |
|   |  Acetaminophen | 200 mg/L  |
|   |  Ibuprofen | 500 mg/L  |
|   |  Theophylline | 100 mg/L  |
|  Urine | Acetaminophen | 3000 mg/L  |
|   |  N-Acetylcysteine | 10 mg/L  |
|   |  Salicyluric acid | 6000 mg/L  |
|   |  Ascorbic acid | 4000 mg/L  |
|   |  Na-Cefoxitin | 12000 mg/L  |
|   |  Gentamycine sulfate | 400 mg/L  |
|   |  Ibuprofen | 4000 mg/L  |

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|  Potential Interferent |   | Highest Concentration Tested Without Significant Interference  |
| --- | --- | --- |
|   | Levodopa | 1000 mg/L  |
|   |  Methyldopa | 2000 mg/L  |
|   |  Ofloxacine | 900 mg/L  |
|   |  Phenazopyridine | 300 mg/L  |
|   |  Tetracycline | 300 mg/L  |

# ONLINE DAT Methadone II

The effects of potential drug interference on the ONLINE DAT Methadone II assay were determined on the cobas pure integrated solutions system using urine samples. Two urine samples containing methadone at positive and negative levels (a final methadone concentration of 225 ng/mL and 375 ng/mL (±25% of the cutoff concentration)) were prepared. In case interference was detected, the interferent was diluted and tested until no more interference was observed. The maximum drug concentration which did not interfere is presented in the table below.

Structurally Unrelated Pharmacological Compounds:

|  Potential Interferent | Drug Concentration (ng/mL) | Methadone Concentration  |   |
| --- | --- | --- | --- |
|   |   |  225 ng/mL (-25% Cutoff, Negative Control) | 375 ng/mL (+25% Cutoff, Positive Control)  |
|  Δ9 THC-9-carboxylic acid | 100,000 | Neg | Pos  |
|  Acetaminophen | 3,000,000 | Neg | Pos  |
|  Acetylsalicylic acid | 100,000 | Neg | Pos  |
|  Aminopyrine | 100,000 | Neg | Pos  |
|  Amitriptyline | 100,000 | Neg | Pos  |
|  Amobarbital | 100,000 | Neg | Pos  |
|  Ampicillin | 100,000 | Neg | Pos  |
|  Ascorbic acid | 4,000,000 | Neg | Pos  |
|  Aspartame | 100,000 | Neg | Pos  |
|  Atropine | 100,000 | Neg | Pos  |
|  Benzocaine | 100,000 | Neg | Pos  |
|  Benzoylecogonine | 100,000 | Neg | Pos  |
|  Benzphetamine | 100,000 | Neg | Pos  |
|  Butabarbital | 100,000 | Neg | Pos  |
|  Caffeine | 100,000 | Neg | Pos  |
|  Calcium hypochlorite | 100,000 | Neg | Pos  |
|  Carbamazepine | 100,000 | Neg | Pos  |
|  Cefoxitin | 12,000,000 | Neg | Pos  |
|  Chlordiazepoxide | 100,000 | Neg | Pos  |
|  Chloroquine | 100,000 | Neg | Pos  |
|  Chlorpheniramine | 100,000 | Neg | Pos  |
|  Cocaine | 100,000 | Neg | Pos  |
|  Codeine | 100,000 | Neg | Pos  |

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K220134 - Page 23 of 34
|  Potential Interferent | Drug Concentration (ng/mL) | Methadone Concentration  |   |
| --- | --- | --- | --- |
|   |   |  225 ng/mL (-25% Cutoff, Negative Control) | 375 ng/mL (+25% Cutoff, Positive Control)  |
|  Cotinine | 100,000 | Neg | Pos  |
|  Cyclobenzaprine | 100,000 | Neg | Pos  |
|  Cyproheptadine | 100,000 | Neg | Pos  |
|  d-Amphetamine | 100,000 | Neg | Pos  |
|  d-Ephedrine | 100,000 | Neg | Pos  |
|  Desipramine | 100,000 | Neg | Pos  |
|  Dextrometorphan | 100,000 | Neg | Pos  |
|  Diazepam | 100,000 | Neg | Pos  |
|  Diphenhydramin | 100,000 | Neg | Pos  |
|  Diphenylhydantoin | 100,000 | Neg | Pos  |
|  Disopyramid | 1,000,000 | Neg | Pos  |
|  dl-Ephedrine | 100,000 | Neg | Pos  |
|  d-Methamphetamine | 100,000 | Neg | Pos  |
|  Dopamine | 100,000 | Neg | Pos  |
|  Doxepin | 100,000 | Neg | Pos  |
|  Doxylamine | 100,000 | Neg | Pos  |
|  d-Phenylpropanolamine | 100,000 | Neg | Pos  |
|  d-Propoxyphen | 100,000 | Neg | Pos  |
|  d-Pseudoephedrine | 100,000 | Neg | Pos  |
|  Ecgonine | 100,000 | Neg | Pos  |
|  Ecgonine methyl ester | 100,000 | Neg | Pos  |
|  EDDP | 100,000 | Neg | Pos  |
|  EMDP | 100,000 | Neg | Pos  |
|  Epinephrine | 100,000 | Neg | Pos  |
|  Erythromycin | 100,000 | Neg | Pos  |
|  Estriol | 100,000 | Neg | Pos  |
|  Fenoprofen | 100,000 | Neg | Pos  |
|  Fluoxetine | 100,000 | Neg | Pos  |
|  Furesemide | 100,000 | Neg | Pos  |
|  Gentamycine sulfate | 400,000 | Neg | Pos  |
|  Gentisic acid | 100,000 | Neg | Pos  |
|  Glutethimide | 100,000 | Neg | Pos  |
|  Guaiacol glycerol ether | 100,000 | Neg | Pos  |
|  Haloperidol | 100,000 | Neg | Pos  |
|  Hydrochlorothiazide | 100,000 | Neg | Pos  |
|  Ibuprofen | 4,000,000 | Neg | Pos  |
|  Imipramine | 100,000 | Neg | Pos  |
|  Isoproterenol | 100,000 | Neg | Pos  |
|  Ketamine | 100,000 | Neg | Pos  |
|  l-Amphetamine | 100,000 | Neg | Pos  |

{23}

K220134 - Page 24 of 34
|  Potential Interferent | Drug Concentration (ng/mL) | Methadone Concentration  |   |
| --- | --- | --- | --- |
|   |  | 225 ng/mL (-25% Cutoff, Negative Control) | 375 ng/mL (+25% Cutoff, Positive Control)  |
|  l-Ephedrine | 100,000 | Neg | Pos  |
|  Levodopa | 1,000,000 | Neg | Pos  |
|  Lidocaine | 100,000 | Neg | Pos  |
|  l-Methamphetamine | 100,000 | Neg | Pos  |
|  l-Norpseudoephedrine | 100,000 | Neg | Pos  |
|  l-Pseudoephedrine | 100,000 | Neg | Pos  |
|  LSD | 100,000 | Neg | Pos  |
|  Maprotiline | 100,000 | Neg | Pos  |
|  MDA | 100,000 | Neg | Pos  |
|  MDMA | 100,000 | Neg | Pos  |
|  Melanin | 10,000 | Neg | Pos  |
|  Meperidin | 100,000 | Neg | Pos  |
|  Methaqualone | 100,000 | Neg | Pos  |
|  Methyldopa | 2,000,000 | Neg | Pos  |
|  Methylphenidate | 100,000 | Neg | Pos  |
|  Mianserin | 100,000 | Neg | Pos  |
|  Morphine | 100,000 | Neg | Pos  |
|  N-acetyl cysteine | 10,000 | Neg | Pos  |
|  Naloxone | 100,000 | Neg | Pos  |
|  Naltrexone | 100,000 | Neg | Pos  |
|  Naproxen | 100,000 | Neg | Pos  |
|  Niacinamide | 100,000 | Neg | Pos  |
|  Nicotine | 100,000 | Neg | Pos  |
|  Nordiazepam | 100,000 | Neg | Pos  |
|  Nordoxepin | 100,000 | Neg | Pos  |
|  Norethindrone | 100,000 | Neg | Pos  |
|  Nortriptyline | 100,000 | Neg | Pos  |
|  Ofloxacin | 90,000 | Neg | Pos  |
|  Orphenadrine | 100,000 | Neg | Pos  |
|  Oxazepam | 100,000 | Neg | Pos  |
|  Penicillin G | 100,000 | Neg | Pos  |
|  Pentobarbital | 100,000 | Neg | Pos  |
|  Perphenazine | 100,000 | Neg | Pos  |
|  Phenazopyridine | 300,000 | Neg | Pos  |
|  Phencyclidine | 100,000 | Neg | Pos  |
|  Phenobarbital | 100,000 | Neg | Pos  |
|  Phenothiazine | 100,000 | Neg | Pos  |
|  Phentermine | 100,000 | Neg | Pos  |
|  Phenylbutazone | 100,000 | Neg | Pos  |
|  Phenylpropanolamine | 100,000 | Neg | Pos  |
|  Procaine | 100,000 | Neg | Pos  |
|  Protriptyline | 100,000 | Neg | Pos  |

{24}

|  Potential Interferent | Drug Concentration (ng/mL) | Methadone Concentration  |   |
| --- | --- | --- | --- |
|   |   |  225 ng/mL (-25% Cutoff, Negative Control) | 375 ng/mL (+25% Cutoff, Positive Control)  |
|  Quetiapine fumarate | 750,000 | Neg | Pos  |
|  Quetiapine Carboxylic Acid | 500,000 | Neg | Pos  |
|  Quetiapine Sulfoxyd | 500,000 | Neg | Pos  |
|  Quinidine | 100,000 | Neg | Pos  |
|  Quinine | 100,000 | Neg | Pos  |
|  Salicyluric acid | 6,000,000 | Neg | Pos  |
|  Secobarbital | 100,000 | Neg | Pos  |
|  β-Phenethylamine | 100,000 | Neg | Pos  |
|  Sulindac | 100,000 | Neg | Pos  |
|  Tetracycline | 300,000 | Neg | Pos  |
|  Tetrahydrozoline | 100,000 | Neg | Pos  |
|  Tramadol | 20,000 | Neg | Pos  |
|  Trifluoperazine | 100,000 | Neg | Pos  |
|  Tyramine | 100,000 | Neg | Pos  |
|  Verapamil | 100,000 | Neg | Pos  |

No interference was found for all tested compounds.

## Elecsys TSH

The effect on the quantitation of the analyte in the presence of potentially interfering drugs was determined by comparing values obtained from samples spiked with 17 commonly and 13 specially used pharmaceutical compounds with the reference sample (unspiked). Two human serum samples (native serum pools) with analyte concentration of approximately 0.3 and 7 µIU/mL were used and tested on the cobas pure integrated solutions system. The summary of results is presented in the table below.

|  Potential Interferent | Highest Concentration Tested Without Significant Interference (mg/L)  |
| --- | --- |
|  Acetylcysteine | 150  |
|  Ampicillin - Na | 75  |
|  Ascorbic acid | 52.5  |
|  Cyclosporine | 1.8  |
|  Cefoxitin | 750  |
|  Heparin | 3300 IU/mL  |
|  Itraconazole | 30  |
|  Levodopa | 7.5  |
|  Methyldopa | 22.5  |
|  Metronidazole | 123  |
|  Phenylbutazone | 321  |
|  Doxycycline | 18  |
|  Acetylsalicylic acid | 30  |
|  Rifampicin | 48  |

K220134 - Page 25 of 34

{25}

|  Potential Interferent | Highest Concentration Tested Without Significant Interference (mg/L)  |
| --- | --- |
|  Acetaminophen | 156  |
|  Ibuprofen | 219  |
|  Theophylline | 60  |
|  Iodide | 0.2  |
|  Levothyroxine | 0.25  |
|  Liothyronine | 0.075  |
|  Carbimazole | 30  |
|  Methimazole | 80  |
|  Propylthiouracil | 300  |
|  Perchlorate | 2000  |
|  Propranolol | 240  |
|  Amiodarone | 200  |
|  Prednisolone | 100  |
|  Hydrocortisone | 200  |
|  Fluocortolone | 100  |
|  Octreotide | 0.3  |

## ONLINE DAT Methadone II Cross Reactivity

The specificity of the ONLINE DAT Methadone II assay for structurally similar compounds was determined by preparing concentration series for each of the compounds listed, using measured signals of the different potential cross reactant concentration to determine the approximate quantity of each compound that is equivalent in assay reactivity to the 300 ng/mL assay cutoff. The percent cross-reactivity was calculated from the ratio of methadone concentration at cutoff level and the calculated cross reactant concentration, that yields the same signal.

Structurally related pharmaceutical compounds:

|  Potential Interferent | Cross reactant units [ng/mL] equivalent to 300 ng/mL methadone | Cross-reactivity (%)  |
| --- | --- | --- |
|  Vortioxetine | 5105 | 5.9  |
|  LUAA34443 | 5216 | 5.8  |
|  Cyamemazine | 8983 | 3.3  |
|  Methotrimeprazine (Levomepromazine) | 9359 | 3.2  |
|  Chlorpromazine | 48833 | 0.6  |
|  Thiothixene | 71795 | 0.4  |
|  Promazine | 184457 | 0.2  |
|  Clomipramine | 251636 | 0.1  |
|  Thioridazine | 264054 | 0.1  |
|  Chlorprothixene | 469530 | 0.1  |
|  Promethazine | 472000 | 0.1  |
|  Trimipramine | 594975 | 0.1  |

K220134 - Page 26 of 34

{26}

4. Assay Reportable Range:

Assays reportable ranges are described in the linearity section (VII.A.2 above).

5. Traceability, Stability, Expected Values (Controls, Calibrators, or Methods):

Traceability

Traceability information for the Glucose HK Gen.3 assay, the ISE indirect Na for Gen.2 test and the Elecsys TSH assay was provided in K191899 and is unchanged.

ONLINE DAT Methadone II assay is traceable to a primary reference method (GC-MS).

On-board Reagent Stability

Glucose HK Gen. 3

A reagent stability study confirmed that the Glucose HK Gen.3 reagent is stable for 26 weeks on-board the cobas pure integrated solutions system.

ONLINE DAT Methadone II

A reagent stability study confirmed that the ONLINE DAT Methadone II reagent is stable for 26 weeks on-board the cobas pure integrated solutions system.

Elecsys TSH

A reagent stability study confirmed that the Elecsys TSH reagent is stable for 16 weeks on-board the cobas pure integrated solutions system.

6. Detection Limit:

The Limit of Blank (LoB), Limit of Detection (LoD) and Limit of Quantitation (LoQ) were determined in accordance with the CLSI EP17-A2 guideline.

The LoB was determined as the 95th percentile of measurements of blank samples. For determination of the LoB for the Glucose HK Gen. 3 assay and the ISE indirect Na for Gen. 2 test, one analyte-free sample was measured with three reagent lots in six runs, distributed over three days, 10 replicates per run, on one cobas pure integrated solutions system. For determination of LoB for the Elecsys TSH assay, five blank samples were measured with three reagent lots in six runs, distributed over three days, two replicates per run, on one cobas pure integrated solutions system. In total, 60 determinations of analyte free samples were obtained. Data analysis was based on determination of the 95th percentile of the 60 measured values. In this design (n=60) the 95th percentile was the average of the 57th and 58th value.

The LoD was determined as the lowest amount of analyte in a sample that can be detected with a 95% probability. For determination of the LoD for the Glucose HK Gen.3 assay, the ISE indirect Na for Gen.2 test and the Elecsys TSH assay, five samples with low analyte content were measured using three reagent lots with two-fold determination per run on one cobas pure integrated solutions system. Six runs distributed over ≥ three days on one instrument were performed. The LoD is defined as the concentration, at which there is a 95% probability that a sample contains analyte. $\mathrm{LoD} = \mathrm{LoB} + 1.653 \times \mathrm{SD}_{\mathrm{total}}$.

K220134 - Page 27 of 34

{27}

The LoQ was determined as the lowest concentration of analyte which can be quantified with a maximum CV of 20% for the Glucose HK Gen.3 assay and the Elecsys TSH assay and a maximum total error (TE) of 30% for the ISE indirect Na for Gen.2 test. For determination of the LoQ for the Glucose HK Gen. 3 assay, the ISE indirect Na for Gen.2 test and the Elecsys TSH assay, five samples with low analyte concentration were measured using three reagent lots on one cobas pure integrated solutions system. For the Glucose HK Gen.3 assay and the Elecsys TSH assay, these samples were tested in one run per day over five days, five replicates per run for each LoQ sample (n=25 per sample). For the ISE indirect Na for Gen.2 test, these samples were tested in two runs per day over three days, two replicates per run for each LoQ sample (n=12 per sample).

The results support the claimed LoB, LoD and LoQ for the Glucose HK Gen.3 assay, the ISE indirect Na for Gen.2 test and the Elecsys TSH assay which are unchanged from the predicate device (K191899) and are summarized in the table below.

|  Assay | Claimed LoB | Claimed LoD | Claimed LoQ  |
| --- | --- | --- | --- |
|  Glucose HK Gen. 3 (serum/plasma, urine, and CSF) | 2 mg/dL (0.11 mmol/L) | 2 mg/dL (0.11 mmol/L) | 2 mg/dL (0.11 mmol/L)  |
|  ISE indirect Na for Gen. 2 (serum, plasma and urine) | 3.5 mmol/L | 4.5 mmol/L | 12.2 mmol/L  |
|  Elecsys TSH (serum) | 0.0025 μIU/mL | 0.005 μIU/mL | 0.005 μIU/mL  |

The upper and lower limits of the measuring range for the Glucose HK Gen.3 assay, the ISE indirect Na for Gen.2 test and the Elecsys TSH assay are supported by the LoQ studies and the linearity studies described above (section VII.A.2).

7. Assay Cut-Off:

Glucose HK Gen. 3, ISE indirect Na for Gen. 2 and Elecsys TSH
Not applicable.

ONLINE DAT Methadone II
Characterization of how the device performs analytically around the claimed cutoff concentration is described in the method comparison section (VII.B.1. below).

8. Accuracy (Instrument):

Not applicable.

9. Carry-Over:

Carry-over information for the Glucose HK Gen.3 assay, the ISE indirect Na for Gen.2 test and the ONLINE DAT Methadone II test is provided in the Method Sheets of these assays. The sponsor has evaluated the special wash program and the carry over evasion rules on the cobas pure integrated solutions system.

K220134 - Page 28 of 34

{28}

B Comparison Studies:

1. Method Comparison with Predicate Device:

Glucose HK Gen. 3

Method comparison studies were performed for serum, urine and CSF using the Glucose HK Gen.3 assay on the cobas pure integrated solutions system and the Glucose HK Gen.3 assay on the cobas pro integrated solutions system (predicate) to assess the differences between the two test systems. Native human samples were tested in singlicate on each test system. Less than 10% of these samples were spiked or diluted to cover the measuring range. The results were analyzed using Passing-Bablok and linear regression analyses. The process was repeated for each application (serum, urine, CSF). A summary of results is presented in the table below.

|  Sample Type | N | Sample Concentration Range (mg/dL) | Passing-Bablok (Slope & Intercept) and Correlation (Kendall tau (t)) | Linear Regression (Slope & Intercept) and Correlation (Pearson (r))  |
| --- | --- | --- | --- | --- |
|  Serum | 69 | 2.2-746.0 | 1.000x + 0.0000
t = 0.991 | 1.000x - 0.0110
r = 1.000  |
|  Urine | 71 | 2.1-742.3 | 1.006x + 0.0007
t = 0.982 | 1.006x + 0.0016
r = 1.000  |
|  CSF | 66 | 5.5-742.3 | 1.000x - 0.0100
t = 0.979 | 0.990x + 0.0202
r = 1.000  |

ISE indirect Na for Gen. 2

Method comparison studies were performed for all sample types using the ISE indirect Na for Gen2. test on the cobas pure integrated solutions system and the ISE indirect Na for Gen2. Test on the cobas pro integrated solutions system (predicate) to assess the differences between the two test systems. Additionally, the results of the candidate test system were compared against flame photometry. Human samples for all sample types were tested in singlicate on each test system. Less than 15% of these samples were spiked or diluted to cover the measuring range. The results were evaluated using Passing-Bablok and linear regression analyses. A summary of results is presented in the table below.

K220134 - Page 29 of 34

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K220134 - Page 30 of 34

|  cobas pure integrated solutions vs. | Sample Type | N | Sample Concentration Range (mmol/L) | Passing-Bablok (Slope & Intercept) and Correlation (Kendall tau (t)) | Linear Regression (Slope & Intercept) and Correlation (Pearson (r))  |
| --- | --- | --- | --- | --- | --- |
|  Cobas pro integrated solutions | Serum | 120 | 81.6-178 | 0.984x + 1.23
t = 0.985 | 0.983x + 1.29
r = 1.000  |
|  Flame photometer | Serum | 120 | 81.5-182 | 1.007x - 1.19
t = 0.942 | 1.001x - 0.258
r = 0.996  |
|  Cobas pro integrated solutions | Plasma | 119 | 84.5-174 | 0.980x + 2.38
t = 0.927 | 0.981x + 2.30
r = 0.999  |
|  Flame photometer | Plasma | 118 | 81.6-176 | 0.985x + 1.38
t = 0.868 | 1.000x - 0.415
r = 0.994  |
|  Cobas pro integrated solutions | Urine | 118 | 23.9-246 | 0.997x + 0.334
t = 0.994 | 1.004x - 0.168
r = 1.000  |
|  Flame photometer | Urine | 105 | 24.9-256 | 0.973x + 1.97
t = 0.975 | 0.976x + 1.80
r = 0.999  |

# ONLINE DAT Methadone II

50 negative urine samples and 50 positive urine samples were evaluated with the ONLINE DAT Methadone II test and methadone concentrations were confirmed by GC-MS. In addition, six samples with methadone concentrations of 75-100 % of the cutoff concentration and 15 samples with methadone concentration of 100-125 % of the cutoff concentration were tested. A summary of results is presented in the tables below.

Qualitative Accuracy Study (cutoff = 300 ng/mL)

|   | Negative samples | GC-MS values (ng/mL)  |   |   |   |
| --- | --- | --- | --- | --- | --- |
|   |   |  Near cutoff |   | Positive samples  |   |
|   |   |  229-256 | 317-370 | 412-1703  |   |
|  cobas pure integrated solutions | + | 0 | 1 | 15 | 50  |
|   |  - | 50 | 5 | 0 | 0  |
|   |  Total | 50 | 6 | 15 | 50  |

Semi-Quantitative Accuracy Study (cutoff = 300 ng/mL)

|   | Negative samples | GC-MS values (ng/mL)  |   |   |   |
| --- | --- | --- | --- | --- | --- |
|   |   |  Near cutoff |   | Positive samples  |   |
|   |   |  229-256 | 317-370 | 412-1703  |   |
|  cobas pure integrated solutions | + | 0 | 1 | 15 | 50  |
|   |  - | 50 | 5 | 0 | 0  |
|   |  Total | 50 | 6 | 15 | 50  |

{30}

K220134 - Page 31 of 34

# Elecsys TSH

A method comparison study was performed using the Elecsys TSH assay on the cobas pure integrated solutions system and the Elecsys TSH assay on the cobas pro integrated solutions system (predicate) to assess the differences between the two test systems. A total of 147 samples (132 native human serum samples, single donors as well as pools and 15 native human serum samples diluted with Diluent Multiassay) were measured in singlicate on each test system. The results were evaluated using Passing-Bablok and linear regression analyses. A summary of results is presented in the table below.

|  Sample Type | N | Sample concentration range (μIU/mL) | Passing-Bablok (Slope & Intercept) and correlation (Kendall tau (t)) | Linear Regression (Slope & Intercept) and correlation Pearson (r)  |
| --- | --- | --- | --- | --- |
|  Serum | 174 | 0.006-98.1 | 0.987x – 0.002
t = 1.00 | 0.983x + 0.016
r = 0.992  |

## 2. Matrix Comparison:

### Glucose HK Gen. 3

To support the use of the Glucose HK Gen.3 assay on the cobas pure integrated solutions system with different sample matrices, a matrix comparison study was conducted by comparing values obtained from samples drawn into serum and plasma collection tubes. Some samples were spiked to cover the measuring range. The result of the serum sample was compared with the result of each plasma sample from the same donor. At least 30 serum/plasma pairs were tested for each type of anticoagulant in one run, in singlicate. The results were evaluated using Passing-Bablok regression analysis. A summary of results is presented in the table below.

|  Comparison | N | Range (mg/dL) | Slope | Intercept | Coefficient r  |
| --- | --- | --- | --- | --- | --- |
|  Serum with gel separation vs. serum | 38 | 3.68-708 | 0.996 | -0.0997 | 1.000  |
|  K_{2}EDTA plasma vs. serum | 50 | 3.68-708 | 0.998 | -0.327 | 1.000  |
|  Li-heparin plasma vs. serum | 50 | 3.68-708 | 1.001 | -1.14 | 0.999  |
|  NaF/K-Oxalate plasma vs. serum | 51 | 3.68-708 | 1.008 | -0.619 | 0.999  |
|  NaF/Na_{2}EDTA plasma vs. serum | 48 | 3.68-708 | 1.005 | 0.334 | 0.999  |
|  NaF/citrate/Na_{2}EDTA plasma vs. serum | 48 | 3.68-708 | 1.003 | -1.39 | 0.999  |
|  KF/Na_{2}EDTA plasma vs. serum vs. | 48 | 3.68-708 | 1.007 | -1.87 | 0.998  |

The resulting data support the method sheet claim that serum tube with separation gel, K2EDTA, Li-heparin, NaF/K-oxalate, NaF/Na2EDTA, NaF/citrate/Na2EDTA, and

{31}

KF/Na₂EDTA plasma tubes are acceptable serum/plasma tubes for use with the Glucose HK Gen.3 assay.

## ISE indirect Na for Gen.2

To support the use of the ISE indirect Na for Gen.2 test on the cobas pure integrated solutions system with other sample matrices, a matrix comparison study was conducted by testing matched Li-heparin plasma and serum samples from the same patients on one cobas pure integrated solutions system. The samples were tested in singlicate. The results were evaluated using Passing-Bablok regression analysis. A summary of results is presented in the table below.

|  Comparison | N | Range (mmol/L) | Slope | Intercept | Coefficient r  |
| --- | --- | --- | --- | --- | --- |
|  Li-heparin plasma vs. Serum | 50 | 85.6-176.3 | 1.003 | -0.885 | 0.999  |

The resulting data support the method sheet claim that serum and Li-heparin plasma tubes are acceptable tubes for use with ISE indirect Na for Gen.2 assay.

## Elecsys TSH

To support the use of the Elecsys TSH assay on the cobas pure integrated solutions system with other sample matrices, a matrix comparison study was conducted by comparing values obtained from samples (native human serum samples) drawn into serum, Li-heparin, K₂EDTA and K₃EDTA plasma tubes. At least 50 serum/plasma pairs per anticoagulant type were tested in singlicate on one cobas pure integrated solutions system. The results were evaluated using Passing Bablok regression analysis. A summary of results is presented in the table below.

|  Comparison | N | Range (μIU/L) | Slope | Intercept | Coefficient r  |
| --- | --- | --- | --- | --- | --- |
|  Li-heparin plasma vs. serum | 51 | 0.008-91.6 | 0.964 | -0.008 | 0.999  |
|  K₂EDTA plasma vs. serum | 52 | 0.008-91.6 | 0.966 | 0.004 | 0.999  |
|  K₃EDTA plasma vs. serum | 52 | 0.008-91.6 | 0.970 | -0.006 | 1.000  |

In addition, a plasma separation tubes (PST) and serum separation tubes (SST) comparison study was conducted, using separation tubes from three separate manufacturers and blood from at least seven donors. Measurements were performed in duplicate using one reagent lot on one cobas pure integrated solutions system and evaluated based on recovery relative to the serum/plasma tube without separating gel (reference).

The results of these studies support the method sheet claim that the following sample types are acceptable to use for Elecsys TSH assay: serum collected using standard serum tubes or tubes containing separating gel, Li-heparin, K₂EDTA, K₃EDTA plasma and plasma tubes containing separating gel.

K220134 - Page 32 of 34

{32}

K220134 - Page 33 of 34

C Clinical Studies:

1. Clinical Sensitivity:
Not applicable.

2. Clinical Specificity:
Not applicable.

3. Other Clinical Supportive Data (When 1. and 2. Are Not Applicable):
Not applicable.

D Clinical Cut-Off:
Not applicable.

E Expected Values/Reference Range:

Expected reference ranges for the assays are as follows:

Glucose HK Gen.3

|  Expected Values  |   |   |
| --- | --- | --- |
|  Sample Type |   | Reference Range  |
|  Serum, plasma | Adults | 74-106 mg/dL  |
|   |  60-90 years | 82-115 mg/dL  |
|   |  > 90 years | 75-121 mg/dL  |
|   |  Children | 60-100 mg/dL  |
|   |  Neonates (1 day) | 40-60 mg/dL  |
|   |  Neonates (> 1 day) | 50-80 mg/dL  |
|  Urine | 24-hour urine | < 2.78 mmol/24 h (<0.5 g/24 h)  |
|   |  Random urine | 1-15 mg/dL  |
|  CSF | Children | 60-80 mg/dL  |
|   |  Adults | 40-70 mg/dL  |

Reference: Tietz NW, ed. Clinical Guide to Laboratory Tests, 4th ed. Philadelphia: WB Saunders Co 2006;444-451.

ISE indirect Na for Gen. 2

|  Expected Values  |   |   |
| --- | --- | --- |
|  Sample Type |   | Reference Range  |
|  Serum, plasma | Adults | 136-145 mmol/L  |
|  Urine | 24 h, Adults | 40-220 mmol/24 h  |

Reference: Tietz NW. Fundamentals of Clinical Chemistry, 5th ed. Burtis CA, Ashwood ER, eds. WB Saunders Co 2001:970, 1004, 1009.

{33}

K220134 - Page 34 of 34

# ONLINE DAT Methadone II

## Qualitative assay

Results of this assay distinguish preliminary positive (≥ 300 ng/mL) from negative samples only. The amount of drug detected in a preliminary positive sample cannot be estimated.

## Semiquantitative assay

Results of this assay yield only approximate cumulative concentrations of the drug and its metabolites.

## Elecsys TSH

|  Expected Values  |   |
| --- | --- |
|  Sample Type | Reference Range  |
|  Serum, plasma | 0.270-4.20 μIU/mL  |

Reference: Ebert C, Bieglmayer C, Igari J, et al. Elecsys TSH, FT4, T4, T-uptake, FT3 and T3. Clinical results of a multicentre study. Wien Klin Wochenschr 1998;110 Suppl 3:27-40.

## F Other Supportive Instrument Performance Characteristics Data:

Not applicable.

## VIII Proposed Labeling:

The labeling supports the finding of substantial equivalence for this device.

## IX Conclusion:

The submitted information in this premarket notification is complete and supports a substantial equivalence decision.

---

**Source:** [https://fda.innolitics.com/submissions/CH/subpart-b%E2%80%94clinical-chemistry-test-systems/CFR/K220134](https://fda.innolitics.com/submissions/CH/subpart-b%E2%80%94clinical-chemistry-test-systems/CFR/K220134)

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